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Literature summary for 1.1.2.B3 extracted from

  • Mennenga, B.; Kay, C.W.; Goerisch, H.
    Quinoprotein ethanol dehydrogenase from Pseudomonas aeruginosa: the unusual disulfide ring formed by adjacent cysteine residues is essential for efficient electron transfer to cytochrome c550 (2009), Arch. Microbiol., 191, 361-367.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
Amine the enzymatic activity of the mutant enzyme in the artificial dye test with N-methylphenazonium methyl sulfate (PMS) and 2,6-dichlorophenol indophenol (DCPIP) at pH 9 does not depend on an activating amine which is essential for wild type activity under these condition Pseudomonas aeruginosa
ethylamine 5 mM, wild-type enzyme shows strict dependence on an activator Pseudomonas aeruginosa

Cloned(Commentary)

Cloned (Comment) Organism
-
Pseudomonas aeruginosa

Protein Variants

Protein Variants Comment Organism
C105A/C106A all pyrroloquinoline quinone-dependent alcohol dehydrogenases contain an unusual disulfide ring formed between adjacent cysteine residues. A mutant enzyme that is lacking this structure is generated by replacing Cys105 and Cys106 with Ala. Heterologously expressed C105A/C106A apoenzyme is successfully converted to enzymatic active holo-enzyme by incorporation of its cofactor pyrroloquinoline quinone (PQQ) in the presence of Ca2+. The enzymatic activity of the mutant enzyme in the artificial dye test with N-methylphenazonium methyl sulfate (PMS) and 2,6-dichlorophenol indophenol (DCPIP) at pH 9 does not depend on an activating amine which is essential for wild type activity under these conditions. The mutant enzyme shows increased Michaelis constants for primary alcohols, while the affinity for the secondary alcohol 2-propanol is unaltered. For all substrates tested the specific activity of the mutant enzyme in the artificial dye test is higher than that found for wild type enzyme. In the ferricyanide test with the natural electron acceptor cytochrome c550 the activity of mutant Cys105Ala/Cys106Ala is 15fold lower than that of wild type enzyme Pseudomonas aeruginosa

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.012
-
ethanol pH 9 in the artificial dye test at fixed concentrations of N-methylphenazonium methyl sulfate and 2,6-dichlorophenol indophenol in the presence of 5 mM ethylamine, wild-type enzyme Pseudomonas aeruginosa
0.062
-
1-propanol pH 9 in the artificial dye test at fixed concentrations of N-methylphenazonium methyl sulfate and 2,6-dichlorophenol indophenol in the presence of 5 mM ethylamine, wild-type enzyme Pseudomonas aeruginosa
0.43
-
1-butanol pH 9 in the artificial dye test at fixed concentrations of N-methylphenazonium methyl sulfate and 2,6-dichlorophenol indophenol in the presence of 5 mM ethylamine, wild-type enzyme Pseudomonas aeruginosa
1.4
-
2-propanol pH 9 in the artificial dye test at fixed concentrations of N-methylphenazonium methyl sulfate and 2,6-dichlorophenol indophenol in the presence of 5 mM ethylamine, wild-type enzyme Pseudomonas aeruginosa
1.5
-
2-propanol pH 9 in the artificial dye test at fixed concentrations of N-methylphenazonium methyl sulfate and 2,6-dichlorophenol indophenol in the presence of 5 mM ethylamine, mutant enzyme C105A/C106A Pseudomonas aeruginosa
3.8
-
1-propanol pH 9 in the artificial dye test at fixed concentrations of N-methylphenazonium methyl sulfate and 2,6-dichlorophenol indophenol in the presence of 5 mM ethylamine, mutant enzyme C105A/C106A Pseudomonas aeruginosa
12
-
ethanol pH 9 in the artificial dye test at fixed concentrations of N-methylphenazonium methyl sulfate and 2,6-dichlorophenol indophenol in the presence of 5 mM ethylamine, mutant enzyme C105A/C106A Pseudomonas aeruginosa
12
-
1-butanol pH 9 in the artificial dye test at fixed concentrations of N-methylphenazonium methyl sulfate and 2,6-dichlorophenol indophenol in the presence of 5 mM ethylamine, mutant enzyme C105A/C106A Pseudomonas aeruginosa
21
-
1,3-Propanediol pH 9 in the artificial dye test at fixed concentrations of N-methylphenazonium methyl sulfate and 2,6-dichlorophenol indophenol in the presence of 5 mM ethylamine, wild-type enzyme Pseudomonas aeruginosa
266
-
1,3-Propanediol pH 9 in the artificial dye test at fixed concentrations of N-methylphenazonium methyl sulfate and 2,6-dichlorophenol indophenol in the presence of 5 mM ethylamine, mutant enzyme C105A/C106A Pseudomonas aeruginosa

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
alcohol + cytochrome c550 Pseudomonas aeruginosa the unusual disulfide ring between the adjacent cysteine residues C105 and C106 is essential for efficient electron transfer at pH 7 from quinoprotein ethanol dehydrogenase to its natural electron acceptor cytochrome c550 aldehyde + reduced cytochrome c550
-
?

Organism

Organism UniProt Comment Textmining
Pseudomonas aeruginosa Q9Z4J7 ATCC17933
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
11
-
substrate: 2-propanol, pH 9 in the artificial dye test at fixed concentrations of N-methylphenazonium methyl sulfate and 2,6-dichlorophenol indophenol in the presence of 5 mM ethylamine, wild-type enzyme Pseudomonas aeruginosa
11
-
substrate: ethanol, pH 9 in the artificial dye test at fixed concentrations of N-methylphenazonium methyl sulfate and 2,6-dichlorophenol indophenol in the presence of 5 mM ethylamine, wild-type enzyme Pseudomonas aeruginosa
15
-
substrate: 1-propanol, pH 9 in the artificial dye test at fixed concentrations of N-methylphenazonium methyl sulfate and 2,6-dichlorophenol indophenol in the presence of 5 mM ethylamine, wild-type enzyme Pseudomonas aeruginosa
20
-
substrate: ethanol, pH 9 in the artificial dye test at fixed concentrations of N-methylphenazonium methyl sulfate and 2,6-dichlorophenol indophenol in the presence of 5 mM ethylamine, mutant enzyme C105A/C106A Pseudomonas aeruginosa
40
-
substrate: 1,3-propanediol, pH 9 in the artificial dye test at fixed concentrations of N-methylphenazonium methyl sulfate and 2,6-dichlorophenol indophenol in the presence of 5 mM ethylamine, wild-type enzyme Pseudomonas aeruginosa
42
-
substrate: 1-butanol, pH 9 in the artificial dye test at fixed concentrations of N-methylphenazonium methyl sulfate and 2,6-dichlorophenol indophenol in the presence of 5 mM ethylamine, wild-type enzyme Pseudomonas aeruginosa
52
-
substrate: 1-propanol, pH 9 in the artificial dye test at fixed concentrations of N-methylphenazonium methyl sulfate and 2,6-dichlorophenol indophenol in the presence of 5 mM ethylamine, mutant enzyme C105A/C106A Pseudomonas aeruginosa
94
-
substrate: 2-propanol, pH 9 in the artificial dye test at fixed concentrations of N-methylphenazonium methyl sulfate and 2,6-dichlorophenol indophenol in the presence of 5 mM ethylamine, mutant enzyme C105A/C106A Pseudomonas aeruginosa
224
-
substrate: 1,3-propanediol, pH 9 in the artificial dye test at fixed concentrations of N-methylphenazonium methyl sulfate and 2,6-dichlorophenol indophenol in the presence of 5 mM ethylamine, mutant enzyme C105A/C106A Pseudomonas aeruginosa
461
-
substrate: 1-butanol, pH 9 in the artificial dye test at fixed concentrations of N-methylphenazonium methyl sulfate and 2,6-dichlorophenol indophenol in the presence of 5 mM ethylamine, mutant enzyme C105A/C106A Pseudomonas aeruginosa

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1,3-propanediol + acceptor artificial dye test at fixed concentrations of N-methylphenazonium methyl sulfate and 2,6-dichlorophenol indophenol Pseudomonas aeruginosa ? + reduced acceptor
-
?
1-butanol + acceptor artificial dye test at fixed concentrations of N-methylphenazonium methyl sulfate and 2,6-dichlorophenol indophenol Pseudomonas aeruginosa butanal + reduced acceptor
-
?
1-propanol + acceptor artificial dye test at fixed concentrations of N-methylphenazonium methyl sulfate and 2,6-dichlorophenol indophenol Pseudomonas aeruginosa propanal + reduced acceptor
-
?
2-propanol + acceptor artificial dye test at fixed concentrations of N-methylphenazonium methyl sulfate and 2,6-dichlorophenol indophenol Pseudomonas aeruginosa 2-propanone + reduced acceptor
-
?
alcohol + cytochrome c550 the unusual disulfide ring between the adjacent cysteine residues C105 and C106 is essential for efficient electron transfer at pH 7 from quinoprotein ethanol dehydrogenase to its natural electron acceptor cytochrome c550 Pseudomonas aeruginosa aldehyde + reduced cytochrome c550
-
?
ethanol + acceptor artificial dye test at fixed concentrations of N-methylphenazonium methyl sulfate and 2,6-dichlorophenol indophenol Pseudomonas aeruginosa acetaldehyde + reduced acceptor
-
?

Synonyms

Synonyms Comment Organism
QEDH
-
Pseudomonas aeruginosa
quinoprotein ethanol dehydrogenase
-
Pseudomonas aeruginosa

Cofactor

Cofactor Comment Organism Structure
pyrroloquinoline quinone
-
Pseudomonas aeruginosa